Publications

[bacteriophages of clostridium histolyticum. isolation and lysotype].		1967	5607844
[on an asporogenic mutant of clostridium histolyticum incapable of synthesizing dipicolinic acid].		1968	5661577
hydrolysis at arginylproline in polypeptides by clostridiopeptidase b.	clostridiopeptidase b, a sulfhydryl protease from clostridium histolyticum, hydrolyzes the arginylproline bond in the synthetic polypeptide methionyllysyl-bradykinin. hydrolysis also occurs at a reduced rate at the lysylarginine bond, further delineating the environment necessary for the minor proteolysis seen with this enzyme at lysine residues. the specificity of clostridiopeptidase b differs from trypsin, which hydrolyzes this synthetic polypeptide only at the lysylarginine bond.	1968	5677534
peptide substrates for a proteinase of clostridium histolyticum.		1968	5722284
complement-inactivating proteinase(s) from clostridium histolyticum.	a proteinase fraction inhibiting the hemolytic activity of guinea pig complement was obtained from supernatant fluids of clostridium histolyticum cultures and purified 150- to 350-fold by ammonium sulfate precipitation, sephadex g-75 gel filtration, and diethylaminoethyl cellulose chromatography. an assay was developed based on the inactivation of hemolytic complement. partially purified anticomplementary preparations were active against casein and were capable of "solubilizing" escherichia coli ...	1968	5724966
toxigenicity of clostridium histolyticum.	nishida, shoki (kanazawa university, kanazawa, japan), and masaaki imaizumi. toxigenicity of clostridium histolyticum. j. bacteriol. 91:477-483. 1966.-from 234 soil samples, 21 strains of clostridium histolyticum of different levels of alpha-toxigenicity were isolated by a new method specially designed for the isolation of this species. the alpha-toxigenicity of freshly isolated strains and of stock strains was closely associated with the potentiality for sporulation, growth, and smooth-colony f ...	1966	5935337
[an anaerobic bacteriophage active on clostridium histolyticum].		1966	5969256
[study of the bacteriophages of clostridium histolyticum strain p].		1967	6062796
[cytologic study of sporulation in clostridium histolyticum. sporogenous strain and sporulation mutants].		1967	6065945
purification and separation of individual collagenases of clostridium histolyticum using red dye ligand chromatography.	six collagenases present in the culture filtrate of clostridium histolyticum have been purified to homogeneity. chromatography over hydroxylapatite, sephacryl s-200, and l-arginine-affi-gel 202 removes the brown pigment and the great majority of the contaminating proteinases active against casein, benzoyl-l-arginine ethyl ester, and elastin. reactive red 120 dye ligand chromatography subdivides the collagenases, which have very similar physicochemical properties, among four fractions. the final ...	1984	6087887
characterization of the individual collagenases from clostridium histolyticum.	the six collagenases (alpha, beta, gamma, delta, epsilon, and zeta) from clostridium histolyticum isolated in the preceding paper [bond, m. d., & van wart, h. e. (1984) biochemistry (first paper of three in this issue)] have been characterized in detail. the molecular weights determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis range from 68 000 to 125 000. isoelectric focusing experiments demonstrate that the isoelectric points of the collagenases are in the 5.35-6.20 range. ...	1984	6087888
relationship between the individual collagenases of clostridium histolyticum: evidence for evolution by gene duplication.	the relationship between the six collagenases (alpha, beta, gamma, delta, epsilon, and zeta) isolated and characterized in the preceding papers [bond, m.d., & van wart, h.e. (1984) biochemistry (preceding two papers in this issue)] has been investigated. chemical modification reactions establish that all six enzymes contain essential carboxyl, tyrosine, and lysine residues. circular dichroism spectra of the peptide bond region show that the secondary structures of the collagenases are very simil ...	1984	6087889
purification and characterization of three forms of collagenase from clostridium histolyticum.	three collagenases from clostridium histolyticum, designated c1, c2, and c3, with apparent molecular weights of 96 000, 92 000, and 76 000 were purified. peptide maps of the enzymes prepared by digestion with staphylococcus aureus v-8 protease were found to be similar. cleavage of native c1 with alpha-chymotrypsin or v-8 protease yielded c2 and c3. this suggested that proteolysis of the mr 96 000 collagenase may have occurred in vivo, producing the other two lower molecular weight enzymes. previ ...	1984	6095892
enhancement of eosinophil effector function by soluble factors released by s. mansoni: role of proteases.	schistosomulum-released products (srp) have been shown to enhance both expression of rat and human eosinophil fc receptors and igg-dependent cytotoxicity. the present work provides additional evidence of the secretion of eosinophil-enhancing factors by schistosomula and other developmental stages of schistosomes, including adult worms. the heat lability, as well as the strong inhibition of the stimulating activity of srp by the protease inhibitor trasylol, suggest that thermolabile proteases sec ...	1983	6190922
[degradation of collagen by the cariogenic bacteria, streptococcus mutans].	the behaviour of cariogenic bacteria (streptococcus mutans) is studied with regard to collagen, which represents 90% of the dentine organic matrix. collagenase activity of cariogenic bacteria is measured with radioactive precursors and gel electrophoresis and compared to reference bacterial collagenase (clostridium histolyticum). labelled collagen substrate has been prepared with two different methods: extraction by 0,5 m acetic acid from young rat skin, previously labelled with l-proline 14c, o ...	1980	6248261
circular dichroism comparative studies of two bacterial collagenases and thermolysin.	the recently isolated and purified collagenase produced by achromobacter iophagus, the collagenase from clostridium histolyticum, and thermolysin, three enzymes having common properties, were studied by circular dichroism. from the spectra of the aqueous solutions obtained in the peptide region, the fraction of alpha helix, beta sheet and aperiodic segments in the three proteins could be estimated. good similarity was found between achromobacter collagenase and thermolysin, which both contain a ...	1980	6250633
[isolation and properties of 3 clostridium histolyticum collagenases].	the collagenases (i, ii and iii) have been obtained in a highly purified state from fresh cultural medium of clostridium histolyticum. the collagenases were similar in their properties to clostridiopeptidase a. the three enzymes differed in their molecular weights, isoelectric points and in some chemical properties. collagenase ii exhibited the most potent hydrolytic activity. its collagenolytic activity was two-fold higher and the peptidase activity was twenty-fold higher as compared with that ...	1980	6252691
enzymatic isolation of cells from bone: cytotoxic enzymes of bacterial collagenase.	the enzymatic isolation of cells from fetal rat calvaria is most effectively achieved with crude clostridium histolyticum collagenase. however, this bacterial collagenase damages the cells during the digestion of the tissue. we have used cell density, as measured by isopycnic centrifugation on polysucrose gradients, as an indicator of cell damage. there are at least two enzymes in crude bacterial collagenase capable of damaging the cells in this tissue. one of these is clostripain that has been ...	1981	6263099
chemical characterization of the homogeneous collagenase from clostridium histolyticum.	pure collagenase (clostridiopeptidase a, ec 3.4.24.3) having a molecular weight of 70 000 was obtained from the culture medium of clostridium histolyticym by a combination of ultrafiltrations, molecular sieve, affinity and hydrophobic chromatography. the value of its specific activity is the highest of those described previously but 6-times lower than that of the collagenase from achromobacter iophagus (ec 3.4.24.8). its amino acid composition differs from previous data, namely by the presence o ...	1981	6266487
a continuous spectrophotometric assay for clostridium histolyticum collagenase.		1981	6269461
identification of essential amino acid residues in clostridium histolyticum collagenase using chemical modification reactions.		1981	6272790
production of collagenase inhibitor by the growth cartilage of embryonic chick bone: isolation and partial characterization.	production of collagenase and collagenase inhibitors by the explants of epiphyseal, metaphyseal and diaphyseal regions of embryonic chick limbs during development has been investigated. collagenase-inhibitory activity was first detected in the culture medium of the diaphyseal region of limbs at stage 36 where cartilage matrix erosion began to occur. however, neither active nor latent collagenase was detected in the media of any regions examined. at stage 38 and later, the maximum production of t ...	1981	6286227
[probable inhibition of a bacterial collagenase by serotonin].	we have previously demonstrated that intraperitoneal injections in rats of collagenase (from clostridium histolyticum) are followed by severe lesions. here we show that the injection of collagenase together with serotonin has no or only small effects. it appears to be partly due to serotonin. creatinin and sulfate which form an injectable complex with serotonin have no influence on this phenomenon. the part of ph 4,4 of the injected solution is discussed. these facts suggest that serotonin may p ...	1982	6290002
bacterial collagenases and their clinical applications.	clostridium histolyticum cultures secrete a specific collagenase which can be precipitated from the medium in relatively large amounts. the enzyme is a metalloproteinase capable of cleaving native collagen types i, ii, iii, iv and v. in addition to being a valuable tool in the laboratory the bacterial collagenase has found clinical applications in the treatment of third degree burns and decubitus, diabetic or arterial ulcers. several cases of successful topical use of the crude enzyme in an oint ...	1982	6295413
inhibition of collagenase from clostridium histolyticum by phosphoric and phosphonic amides.	di- and tripeptides with sequences present in collagen that are known to occupy the s1' through s3' subsites at the active site of the collagenase from clostridium histolyticum do not themselves inhibit this zinc protease. thus glycylproline, glycylprolylalanine, and their c-terminal amides are not inhibitors. n alpha-phosphorylglycylproline, n alpha-phosphorylglycyl-l-prolyl-l-alanine, and their c-terminal amides are weak inhibitors with ic50's (concentration causing half-maximal inhibition) of ...	1983	6313042
characterization of the metalloproteinase inhibitor produced by bovine articular chondrocyte cultures.	primary cultures of bovine articular chondrocytes release a latent metalloproteinase which is activated by incubation with organomercurials to degrade proteoglycans. all the enzyme present in the culture medium is latent and binds to columns of heparin-sepharose. the yield of activity from the heparin-sepharose columns (measured after organomercurial treatment) is approximately 300-1000% depending on the chondrocyte culture batch. recombination of column fractions shows that the increase in acti ...	1983	6313063
enzymatic isolation of cells from neonatal calvaria using two purified enzymes from clostridium histolyticum.	the enzymatic isolation of cells with bacterial collagenase has proved to be a powerful technique for the study of a wide variety of tissues. unfortunately, for some applications such as the isolation of cells from membranous bone, the cellular damage that results from the exposure of the cells to cytotoxic contaminants of bacterial collagenase has limited the usefulness of this approach. the use of chromatographically purified collagenase alone is often ineffective or very slow to release cells ...	1983	6315459
degradation of basement membrane collagen by proteinases from human gingiva, leukocytes and bacterial plaque.	basement membranes in human gingiva are found in dento-epithelial junction, epithelial-connective tissue junction and endothelium-connective-tissue junction where they have important attachment and filtering functions. the ability of plaque and gingiva-derived proteinases to degrade type iv collagen, the major protein of basement membranes, was examined in vitro. the basement membrane collagen (type iv) isolated from bovine lens capsules was incubated in the presence of enzyme samples. the degra ...	1983	6315910
characterization of a large fragment from annelid cuticle collagen and its relationship to the intact molecule.	digestion of the cuticle collagen from the annelid nereis virens with clostridium histolyticum collagenase yields a native, collagenase-resistant fragment (ccrf) of the molecule with an mr of about 900,000 (kimura and tanzer, j. biol. chem. 252: 8018, 1977). we have produced 940 nm long, sls-crystallites from the collagenase-resistant fragment; the sls pattern matches a region at one end of the 2,400 nm sls obtained from intact cuticle collagen. upon denaturation, the fragment yields two subunit ...	1983	6321095
isolation and ultrastructure of rabbit ovarian mesothelium (surface epithelium).	this study evaluated the efficacy of various dissociating procedures for isolating mesothelial or surface epithelial cells from rabbit ovaries. the procedure which provided the highest and most homogenous cell yield involved the following sequence: (a) incubation of whole ovaries for 60 min in clostridium histolyticum collagenase, type i (300 u/ml); (b) vortexing of ovaries at 30 rpm for 60 s in medium 199; (c) removal of partially detached surface epithelium by gentle scraping with a microdisse ...	1984	6392121
[neutral and basic compounds present in gases produced by clostridium histolyticum, clostridium hastiforme and clostridium ghoni cultured under vacuum in sodium thioglycolate glucose broth].	the study of gases produced by clostridium histolyticum, c. hastiforme and c. ghoni cultured under vacuum in sodium thioglycolate glucose broth makes it possible to detect compounds not described before for these bacteria, and confirms the production of methane in anaerobic spore-forming bacteria.	1982	6817857
oxygen-dependent metronidazole-resistance of clostridium histolyticum.	sensitivity determinations with 6 strains of clostridium histolyticum showed that the inhibitory action of metronidazole was highly dependent on the oxygen concentration of the environment. under anaerobic conditions they were sensitive but at increased oxygen concentrations moderately sensitive or resistant. the flexible resistance of this aerotolerant anaerobe against metronidazole may interfere with results of sensitivity determinations, estimation of blood levels and it may influence the eff ...	1981	7269847
augmentation of permeability in the bronchial epithelium by the house dust mite allergen der p1.	sequence analyses have revealed the existence of homology between certain aeroallergens and proteolytic enzymes. this homology can be expressed functionally, but its significance to airway pathophysiology is unknown. studies with madin-darby canine kidney cells and canine tracheal epithelial cells grown on plastic substrata or matrix proteins suggest that der p1, a major allergen from the house dust mite dermatophagoides pteronyssinus and a cysteine proteinase, or the unfractionated growth mediu ...	1995	7695916
different roles of class i and class ii clostridium histolyticum collagenase in rat pancreatic islet isolation.	crude clostridium histolyticum collagenase was purified by gel filtration and fractionated by anion exchange chromatography into class i with high collagen digestion activity (cda) and low falgpa (2-furanacryloyl-l-leucylglycyl-l-prolyl-l-alanine) hydrolysis activity (fha), class ii with low cda and high fha, and a fraction called class i/ii with intermediate activities. the roles of these collagenase classes in rat pancreatic islet isolation were investigated. dissociations were carried out wit ...	1995	7859945
mediation by bradykinin of rat paw oedema induced by collagenase from clostridium histolyticum.	1. collagenases are thought to play a major role in the pathology of gas gangrene caused by clostridium histolyticum, because they can destroy the connective tissue barriers. we investigated possible mediators involved in the oedema formation and plasma protein extravasation which follow the injection of a collagenase (ec 3.4.24.3) from clostridium histolyticum into one hind paw of anaesthetized rats. 2. the magnitude of the oedema following a subplantar injection was dependent on the dose of co ...	1994	7915609
[proteinases with various substrate specificities in structural studies of yeast cell walls].	different proteins are revealed in cell wall of yeast cells candida utilis by means of specific proteolysis with subtilisins tv and 72, trypsin and purified collagenase of clostridium histolyticum. some of them were characterized by resistance to trypsin and sensitivity to subtilisin tv. in young cells this group is represented essentially by a protein of 33 kd, which appears to be one of the structural proteins, binding fibrillae of carbohydrate. other proteins proved to be sensitive to both tr ...	1994	7945458
cell-associated collagenolytic activity by group b streptococci.	group b streptococci (gbs) are important pathogens in neonatal sepsis, pneumonia, and meningitis. the ability of gbs to invade the collagen-rich amniotic membrane of the placenta has been shown in vitro. in the presence of gbs, the collagen fibrils of the amnion appear disordered, suggesting a role for gbs in premature rupture of membranes. sodium dodecyl sulfate-polyacrylamide gel electrophoresis, sephadex g-200 column chromatography, and gelatin zymograms were used in this study to characteriz ...	1994	7960147
cloning and nucleotide sequence analysis of the colh gene from clostridium histolyticum encoding a collagenase and a gelatinase.	the colh gene encoding a collagenase was cloned from clostridium histolyticum jcm 1403. nucleotide sequencing showed a major open reading frame encoding a 116-kda protein of 1,021 amino acid residues. the deduced amino acid sequence contains a putative signal sequence and a zinc metalloprotease consensus sequence, hexxh. a 116-kda collagenase and a 98-kda gelatinase were copurified from culture supernatants of c. histolyticum. while the former degraded both native and denatured collagen, the lat ...	1994	7961400
heterologous expression of the clostripain gene from clostridium histolyticum in escherichia coli and bacillus subtilis: maturation of the clostripain precursor is coupled with self-activation.	clostripain-specific antibodies were used to analyse the maturation of clostripain prepro-enzyme and core protein heterologously synthesized in escherichia coli and bacillus subtilis. core protein purified from e. coli cells harbouring plasmid phm3-23 underwent calcium-dependent, self-triggered maturation. concomitantly, the inactive form of the enzyme was converted into an active form, demonstrating the self-activation capacity of the clostripain core protein. as judged from western blot analys ...	1994	8025682
the specificity of clostripain from clostridium histolyticum. mapping the s' subsites via acyl transfer to amino acid amides and peptides.	the s' subsite specificity of clostripain from clostridium histolyticum was investigated by acyl transfer to libraries of amino acid amides, ala-xaa dipeptides, proline derivatives and pentapeptides using n alpha-benzoyl-l-arginine ethyl ester as acyl donor. a pentapeptide library consisting of 29 pentapeptides with general structure xaa-ala-ala-ala-gly, ala-xaa-ala-ala-gly and ala-ala-xaa-ala-gly, where xaa represents gly, ala, pro, leu, phe, asp, glu, arg and lys, was prepared by solid-phase s ...	1994	8055964
[interaction of collagenases with certain peptide substrates and inhibitors].	interactions of collagenases i and ii (clostridiopeptidases) from clostridium histolyticum with hexapeptide substrates in which some l-proline residues are replaced by their d-analogues, as well as with the tripeptide chloromethyl ketone z-gly-pro-gly-ch2cl were studied. a role of stereochemistry of the amino acid residues in the substrate was established and differences between the collagenases, with regard to their specific requirements to substrates, were revealed. the tripeptide chloromethyl ...	1994	8166757
collagen degradation by three clostridium histolyticum collagenase fractions with different substrate specificities.		1994	8171590
synthesis of n alpha-[3h]acetyl-l-lysine chloromethyl ketone and its use in the fluorographic detection of proteases.	tritiated n alpha-acetyl-l-lysine chloromethyl ketone (alck) was synthesized on a laboratory scale for use as an active-site-directed affinity label in the fluorographic detection of proteases after sodium dodecyl sulfate-polyacrylamide gel electrophoresis (sds-page). the synthesis involved acetylation of n epsilon-benzyloxycarbonyl-l-lysine chloromethyl ketone with [3h]acetic anhydride just before the removal of the benzyloxycarbonyl group. by this method, [3h]alck with a specific activity of 2 ...	1993	8250226
the heterodimeric protease clostripain from clostridium histolyticum is encoded by a single gene.	clostripain (ec 3.4.22.8) is a heterodimeric cysteine endopeptidase with strict specificity for arg-xaa peptidyl bonds. it is secreted by clostridium histolyticum strains. for the first time we present evidence that both polypeptide chains of native clostripain are encoded by a single gene. dna sequencing of two overlapping genomic dna fragments revealed a single open reading frame (orf) of 1581 nucleotides encoding a polypeptide of 526 amino acid residues. the orf is preceded by canonical trans ...	1993	8341259
both medullasin and human leukocyte elastase are essentially devoid of elastinolytic activity.	elastinolytic activity of medullasin was investigated precisely and compared with that of human leukocyte elastase, because the structure of medullasin is quite similar to that of human neutrophil elastase, which was reported to have elastinolytic activity. when elastinolytic activity of medullasin and human leukocyte elastase was determined by employing unstained elastin fibers and measuring the increase in 280-nm absorbance of the supernatant, elastinolytic activity amounting to several percen ...	1993	8407864
a simple in vitro method for evaluating the efficacy of different batches of crude clostridium histolyticum collagenase for islet isolation.		1995	8539957
university of wisconsin solution inhibits the class ii collagenase within crude clostridium histolyticum collagenase.		1995	8539958
recombinant enzymes for islet isolation: purification of a collagenase from clostridium histolyticum and cloning/expression of the gene.		1995	8539959
active site structure of a hemagglutinating protease from porphyromonas gingivalis: similarity to clostripain.	the active site of a 44-kda hemagglutinating arginine-specific protease from the putative periodontopathogen porphyromonas gingivalis was specifically labeled with n alpha-[3h]acetyllysine chloromethyl ketone. after enzymatic digestion of the labeled enzyme, a labeled active site peptide was isolated by hplc. the sequence of the active site peptide was determined, after its treatment with nabh4 to reduce the ketone group of the reagent moiety, to be asp-val-ala-cys-val-asn-gly. the cysteine resi ...	1995	8595395
demonstration that 1-trans-epoxysuccinyl-l-leucylamido-(4-guanidino) butane (e-64) is one of the most effective low mr inhibitors of trypsin-catalysed hydrolysis. characterization by kinetic analysis and by energy minimization and molecular dynamics simulation of the e-64-beta-trypsin complex.	1-trans-epoxysuccinyl-l-leucylamido(4-guanidino)butane (e-64) was shown to inhibit beta-trypsin by a reversible competitive mechanism; this contrasts with the widely held view that e-64 is a class-specific inhibitor of the cysteine proteinases and reports in the literature that it does not inhibit a number of other enzymes including, notably, trypsin. the k1, value (3 x 10(-5) m) determined by kinetic analysis of the hydrolysis of n alpha-benzoyl-l-arginine 4-nitroanilide in tris/hcl buffer, ph ...	1996	8670152
clostripain linker deletion variants yield active enzyme in escherichia coli: a possible function of the linker peptide as intramolecular inhibitor of clostripain automaturation.	the clostripain core protein is composed of the light and heavy chain subunits linked by a nonapeptide into a single polypeptide chain [mol. gen. genet. 240: 140, 1993]. linker removal is due to autocatalytic processing yielding active heterodimeric enzyme. we have expressed mutationally altered core protein variants in the heterologous host escherichia coli to gain further insight into the process of clostripain automaturation. in a mutationally created cys231 --> ser variant, heterodimer forma ...	1996	8875906
fractions from commercial collagenase preparations: use in enzymic isolation of the islets of langerhans from porcine pancreas.	transplantation of isolated islets of langerhans is an intriguing possibility for the treatment of diabetes mellitus. the isolation of islets from pancreata requires the specific dissociation of the tissue. commercial collagenases from clostridium histolyticum are widely used for this purpose. unfortunately, the effectiveness of these commercial enzymes is not predictable and differs considerably between suppliers and even from lot to lot. this is due mainly to differences in their specific coll ...	1996	8889213
identification and analysis of a collagenolytic activity in streptococcus mutans.	streptococcus mutans is an important pathogen in coronal caries and is implicated in dental root decay by its ability to bind collagen from various sources. in the present study, electron microscopic analysis demonstrated the ability of s. mutans to bind and to disrupt collagen fibrils of the amniotic membrane. the synthetic peptide falgpa, which is similar in structure to collagen, was degraded by s. mutans, with a lower level of falgpa hydrolytic activity observed in sucrose-grown cells compar ...	1997	8939802
[collagenolytic enzymes synthesized by microorganisms].	the review uses data obtained by the authors and available in the literature to discuss microbial collagenolytic enzymes, widely employed in scientific research, biotechnology, and medicine. collagenases differing in their structure and the specificity of their action on collagen fibrils were isolated from bacteria (including marine isolates), actinomycetes, and fungi. collagenases produced by clostridium histolyticum and achromobacter iophagus are the best studied enzymes; both are metalloenzym ...	1996	8992239
expression of the colh gene encoding clostridium histolyticum collagenase in bacillus subtilis and its application to enzyme purification.	the colh gene encoding 116-kda collagenase of clostridium histolyticum (ccolh) was cloned into an escherichia coli-bacillus subtilis shuttle vector to develop a method for purification of recombinant collagenase (rcolh). when plasmid pjcm310 containing the colh gene was introduced into b. subtilis db104 and the transformant was grown in lb broth at 37 c, stability of the plasmid was not maintained. however, stability was partly improved by growing the transformant in a modified lb broth containi ...	1996	9013490
involvement of 5-hydroxytryptamine and bradykinin in the hyperalgesia induced in rats by collagenase from clostridium histolyticum.	the involvement of bradykinin, 5-hydroxytryptamine, substance p and prostanoids in the hyperalgesia elicited by collagenase in rat paw was investigated. collagenase (100 micrograms) induced a slight hyperalgesia in kininogen deficient rats in comparison with the behavioural response obtained in normal rats. lisinopril (10(-5) m), and angiotensin-converting enzyme inhibitor, increased the duration of the hyperalgesia elicited in normal rats. ondansetron (0.5 to 5 mumol/kg), a 5-ht3 antagonist, su ...	1997	9151293
a trypanosoma cruzi-secreted 80 kda proteinase with specificity for human collagen types i and iv.	specific interactions between parasites and extracellular matrix components are an important mechanism in the dissemination of chagas' disease. binding of the extracellular matrix proteins to trypanosoma cruzi receptors has been described as a significant step in this phenomenon. in this study, a specific proteinase activity was identified in cell-free extracts of amastigote, trypomastigote and epimastigote forms of t. cruzi using the collagenase fluorogenic substrate n-suc-gly-pro-leu-gly-pro-7 ...	1997	9224638
histochemical analysis of the role of class i and class ii clostridium histolyticum collagenase in the degradation of rat pancreatic extracellular matrix for islet isolation.	to understand why class ii clostridium histolyticum collagenase is much more effective than class i in the isolation of rat pancreatic islets, we analyzed the role of these collagenases in pancreatic tissue dissociation. crude collagenase was purified and then fractionated into class i and ii with different enzyme activities and protein compositions. pancreatic tissue was incubated with either class i, class ii, or class i + ii, with or without added protease, under conditions that eliminated en ...	1997	9258514
a study of the collagen-binding domain of a 116-kda clostridium histolyticum collagenase.	the clostridium histolyticum 116-kda collagenase consists of four segments, s1, s2a, s2b, and s3. a 98-kda gelatinase, which can degrade denatured but not native collagen, lacks the c-terminal fragment containing a part of s2b and s3. in this paper we have investigated the function of the c-terminal segments using recombinant proteins. full-length collagenase degraded both native type i collagen and a synthetic substrate, pz-peptide, while an 88-kda protein containing only s1 and s2a (s1s2a) deg ...	1998	9452493
collagen-binding growth factors: production and characterization of functional fusion proteins having a collagen-binding domain.	the autocrine/paracrine peptide signaling molecules such as growth factors have many promising biologic activities for clinical applications. however, one cannot expect specific therapeutic effects of the factors administered by ordinary drug delivery systems as they have limited target specificity and short half-lives in vivo. to overcome the difficulties in using growth factors as therapeutic agents, we have produced fusion proteins consisting of growth factor moieties and a collagen-binding d ...	1998	9618531
variations of bacterial populations in human feces measured by fluorescent in situ hybridization with group-specific 16s rrna-targeted oligonucleotide probes.	six 16s rrna-targeted oligonucleotide probes were designed, validated, and used to quantify predominant groups of anaerobic bacteria in human fecal samples. a set of two probes was specific for species of the bacteroides fragilis group and the species bacteroides distasonis. two others were designed to detect species of the clostridium histolyticum and the clostridium lituseburense groups. another probe was designed for the genera streptococcus and lactococcus, and the final probe was designed f ...	1998	9726880
gene duplication and multiplicity of collagenases in clostridium histolyticum.	clostridium histolyticum collagenase contains a number of different active components. previously we have shown that colh encodes a 116-kda collagenase (colh) and a 98-kda gelatinase. we purified a different 116-kda collagenase (colg) from the culture supernatant and sequenced its gene (colg). we also identified four other gelatinases (105, 82, 78, and 67 kda) and determined their n-terminal amino acid sequences, all of which coincided with that of either colg or colh. hybridization experiments ...	1999	9922257
identification of metal ligands in the clostridium histolyticum colh collagenase.	a clostridium histolyticum 116-kda collagenase has an h415exxh motif but not the third zinc ligand, as found in already characterized zinc metalloproteinases. to identify its catalytic site, we mutated the codons corresponding to the three conserved residues in the motif to other amino acid residues. the mutation affecting his415 or his419 abolished catalytic activity and zinc binding, while that affecting glu416 did the former but not the latter. these results suggest that the motif forms the c ...	1999	10217773
imaging real-time proteolysis of single collagen i molecules with an atomic force microscope.	the dynamic process of synthesis and degradation of extracellular matrix molecules, including various collagens, is important in normal physiological functions and pathological conditions. existing models of collagen enzymatic degradation reactions are derived from bulk biochemical assays. in this study, we have imaged in real-time individual collagen i molecules and their proteolysis by clostridium histolyticum collagenases in phosphate-buffered saline (pbs) with atomic force microscopy (afm). ...	1999	10433702
the 105-kda protein component of bacillus cereus non-haemolytic enterotoxin (nhe) is a metalloprotease with gelatinolytic and collagenolytic activity.	a sequence of 91 amino acids residues, probably starting from the n-terminal of the mature protein, was determined for the 105-kda protein of the non-haemolytic enterotoxin of bacillus cereus. the last part of this sequence was similar to parts of the n-terminal portions of two collagenases of clostridium histolyticum and clostridium perfringens. zymography, with intact collagen fibril and gelatin as substrates, showed that the 105-kda protein had collagenolytic and gelatinolytic activity. the 1 ...	1999	10499286
protease inhibitors. part 7. inhibition of clostridium histolyticum collagenase with sulfonylated derivatives of l-valine hydroxamate.	sulfonylated l-valine hydroxamate derivatives were obtained by reaction of alkyl/arylsulfonyl halides with the title amino acid, followed by treatment with benzyl chloride, and conversion of the cooh moiety to the conhoh group. other derivatives were obtained by reaction of n-benzyl-l-valine with arylisocyanates, arylsulfonylisocyanates or benzoylisothiocyanate, followed by the similar conversion of the cooh into the conhoh moiety, with hydroxylamine in the presence of carbodiimides. the obtaine ...	2000	10699384
protease inhibitors. part 8: synthesis of potent clostridium histolyticum collagenase inhibitors incorporating sulfonylated l-alanine hydroxamate moieties.	a series of hydroxamates was prepared by reaction of alkyl/arylsulfonyl halides with n-2-chlorobenzyl-l-alanine, followed by conversion of the cooh moiety to the conhoh group, with hydroxylamine in the presence of carbodiimides. other structurally related compounds were obtained by reaction of n-2-chlorobenzyl-l-alanine with aryl isocyanates, arylsulfonyl isocyanates or benzoyl isothiocyanate, followed by the similar conversion of the cooh into the conhoh moiety. the new compounds were assayed a ...	2000	10732980
protease inhibitors: synthesis of clostridium histolyticum collagenase inhibitors incorporating sulfonyl-l-alanine hydroxamate moieties.	a series of hydroxamates was obtained by the reaction of n-(4-nitrobenzyl)-l-alanine with alkyl/arylsulfonyl halides, followed by conversion of the cooh group into conhoh. structurally-related compounds were prepared similarly by using arylsulfonyl isocyanates, aryl isocyanates or arylsulfenyl halides instead of the sulfonyl halides. many of the new compounds showed nanomolar affinity for the bacterial collagenase isolated from the pathogen clostridium histolyticum.	2000	10743957
protease inhibitors - part 5. alkyl/arylsulfonyl- and arylsulfonylureido-/arylureido- glycine hydroxamate inhibitors of clostridium histolyticum collagenase.	reaction of alkyl/arylsulfonyl halides with glycine afforded a series of derivatives which were first n-benzylated by treatment with benzyl chloride, and then converted to the corresponding hydroxamic acids with hydroxylamine in the presence of carbodiimide derivatives. other derivatives were obtained by reaction of n-benzyl-glycine with aryl isocyanates, arylsulfonyl isocyanates or benzoyl isothiocyanate, followed by conversion of their cooh group into the conhoh moiety, as mentioned above. the ...	2000	10785556
protease inhibitors: synthesis of potent bacterial collagenase and matrix metalloproteinase inhibitors incorporating n-4-nitrobenzylsulfonylglycine hydroxamate moieties.	a series of compounds was prepared by reaction of alkyl/arylsulfonyl halides with n-4-nitrobenzylglycine, followed by conversion of the cooh to the conhoh group, with hydroxylamine in the presence of carbodiimides. other structurally related compounds were obtained by reaction of n-4-nitrobenzylglycine with aryl isocyanates, arylsulfonyl isocyanates, or benzoyl isothiocyanate, followed by the similar conversion of the cooh into the conhoh moiety. another subseries of derivatives was prepared fro ...	2000	10794702
protease inhibitors. part 12. synthesis of potent matrix metalloproteinase and bacterial collagenase inhibitors incorporating sulfonylated n-4-nitrobenzyl-beta-alanine hydroxamate moieties.	n-4-nitrobenzyl-beta-alanine was reacted with alkyl/arylsulfonyl halides, followed by conversion of the cooh to the conhoh group. structurally related compounds were obtained by reaction of n-4-nitrobenzyl-beta-alanine with aryl isocyanates, arylsulfonyl isocyanates or benzoyl isothiocyanate, followed by similar conversion of the cooh into the conhoh moiety. another subseries of derivatives was prepared from sulfanilyl- or metanilyl-4-nitrobenzyl-beta-alanine by reaction with arylsulfonyl isocya ...	2000	10913755
protease inhibitors: synthesis of l-alanine hydroxamate sulfonylated derivatives as inhibitors of clostridium histolyticum collagenase.	l-alanine hydroxamate derivatives were obtained by reaction of alkyl/arylsulfonyl halides with l-alanine, followed by treatment with benzyl chloride, and conversion of the cooh moiety to the conhoh group with hydroxylamine in the presence of carbodiimides. other derivatives were obtained by reaction of n-benzyl-alanine with aryl isocyanates, arylsulfonyl isocyanates or benzoyl isothiocyanate, followed by a similar conversion of the cooh to the conhoh moiety. the obtained compounds were assayed a ...	2000	10938538
carbonic anhydrase and matrix metalloproteinase inhibitors: sulfonylated amino acid hydroxamates with mmp inhibitory properties act as efficient inhibitors of ca isozymes i, ii, and iv, and n-hydroxysulfonamides inhibit both these zinc enzymes.	the 14 different carbonic anhydrase (ca, ec 4.2.1.1) isozymes as well as the 23 different matrix metalloproteinases (mmps) isolated up to now in higher vertebrates play important physiological functions in these organisms. unsubstituted sulfonamides act as high-affinity inhibitors for the first type of these enzymes, whereas hydroxamates strongly inhibit the latter ones. since the active site geometry around the zinc ion in these two types of metalloenzymes is rather similar, we tested whether s ...	2000	11020282
effects of the non-peptide b2 receptor antagonist fr173657 in models of visceral and cutaneous inflammation.	the non-peptide b2 receptor antagonist (e)-3-(6-acetamido-3-pyridyl)-n-[n-[2,4-dichloro-3-[(2-methyl-8-quinolin yl)oxymethyl]phenyl]-n-methylaminocarbonylmethyl]acrylamide (fr173657) was compared to the peptide antagonist icatibant in models of visceral and cutaneous inflammation.	2000	11089906
atomic force microscopy of intact and digested collagen molecules.	the present study was performed to analyse the structure of non-digested and digested collagen type i molecules by atomic force microscopy (afm). collagen type i molecules from the bovine skin were diluted with 0.05 n acetic acid, spread on a mica plate, air-dried and observed by non-contact mode afm in air. collagen molecules digested with clostridium histolyticum collagenase were also examined by afm. intact collagen type i molecules were observed as twisted threads ranging mainly between 280 ...	2000	11108030
substrate recognition by the collagen-binding domain of clostridium histolyticum class i collagenase.	clostridium histolyticum type i collagenase (colg) has a segmental structure, s1+s2+s3a+s3b. s3a and s3b bound to insoluble collagen, but s2 did not, thus indicating that s3 forms a collagen-binding domain (cbd). because s3a+s3b showed the most efficient binding to substrate, cooperative binding by both domains was suggested for the enzyme. monomeric (s3b) and tandem (s3a+s3b) cbds bound to atelocollagen, which contains only the collagenous region. however, they did not bind to telopeptides immo ...	2001	11121400
metal content and biochemical analyses of a recombinant collagenase prtv from vibrio parahaemolyticus.	prtv is an extracellular metalloprotease of vibrio parahaemolyticus and regarded as a collagenase. inductively coupled plasma-optical emission spectrometry analysis indicated that the recombinant prtv contains 1 mol of zinc per mol of the native enzyme. on the basis of a kinetic study using 2-furanacryloyl-leu-gly-pro-ala (falgpa, the specific substrate for bacterial collagenase) as a substrate, it was suggested that metal ions may play a significant role in the binding and catalytic steps of th ...	2000	11128063
infective endocarditis due to clostridium histolyticum.		2000	11168053
salivary histatin 5 is a potent competitive inhibitor of the cysteine proteinase clostripain.	histatin 5 is a low molecular weight salivary protein which is known to exhibit inhibitory activity against several proteinases, including the cysteine proteinases gingipains. the purpose of this study was to characterize the effect of salivary histatin on the proteolytic activity of the cysteine proteinase clostripain derived from the pathogen clostridium histolyticum. using a synthetic nitroanilide substrate, we studied in detail the inhibition of clostripain by histatin 5 and compared the eff ...	2001	11231021
protease inhibitors: synthesis of a series of bacterial collagenase inhibitors of the sulfonyl amino acyl hydroxamate type.	a series of sulfonyl amino acyl hydroxamates incorporating alkyl/arylsulfonyl-n-2-nitrobenzyl-l-alanine was prepared. related compounds were obtained by reaction of n-2-nitrobenzyl-l-ala with aryl isocyanates, arylsulfonyl isocyanates, or benzoyl isothiocyanate, followed by the conversion of the cooh into the conhoh moiety. the new compounds were assayed as inhibitors of the clostridium histolyticum collagenase (chc), a bacterial protease involved in the degradation of extracellular matrix. many ...	2001	11405662
oligofructose and long-chain inulin: influence on the gut microbial ecology of rats associated with a human faecal flora.	dietary incorporation of fermentable, indigestible fructans may be of benefit to gastrointestinal health by providing short-chain fatty acids, stimulating the proliferation of bifidobacteria or lactobacilli and suppressing potential pathogenic organisms in the gut. we tested the hypothesis that the effects of fructans on caecal, colonic and faecal short-chain fatty acid concentration and microflora composition depend on their chain length. germ-free rats associated with a human faecal flora were ...	2001	11502244
a thermostable collagenolytic protease with a very large molecular mass produced by thermophilic bacillus sp. strain mo-1.	a collagenolytic protease was purified to homogeneity from thermophilic bacillus sp. strain mo-1. the protease from strain mo-1 showed high activity toward type i and iv collagens and gelatin. however, peptide substrates (4-phenylazobenzyloxycarbonyl-pro-leu-gly-pro-arg and 2-furylacryloyl-leu-gly-pro-ala) for collagenases were inert as substrates. the collagenolytic protease cleaved oxidized insulin b-chain at 11 sites and degraded type i and iv collagens into anonymous small pieces, suggesting ...	2001	11693905
rapid determination of substrate specificity of clostridium histolyticum beta-collagenase using an immobilized peptide library.	the molecular basis of the substrate specificity of clostridium histolyticum beta-collagenase was investigated using a combinatorial method. an immobilized positional peptide library, which contains 24,000 sequences, was constructed with a 7-hydroxycoumarin-4-propanoyl (cop) fluorescent group attached at the n terminus of each sequence. this immobilized peptide library was incubated with c. histolyticum beta-collagenase, releasing fluorogenic fragments in the solution phase. the relative substra ...	2002	11724807
characterization of an important enzymatic component in collagenase that is essential for the effective digestion of the human and porcine pancreas.	recent clinical results from edmonton have demonstrated the feasibility of achieving normoglycemia in type i diabetic patients by islet transplantation. one of the key issues in obtaining this success was transplanting sufficient numbers of islets by sequential transplants. although the development of semipurified endotoxin-free clostridium histolyticum-derived collagenase (liberase) has improved islet yields from the human pancreas, batch-to-batch variation and loss of activity with time still ...	2001	11814113
collagen-binding domain of a clostridium histolyticum collagenase exhibits a broad substrate spectrum both in vitro and in vivo.	the substrate spectrum of the tandem collagen-binding domain (cbd) of clostridium histolyticumclass i collagenase (colg) was examined both in vitro and in vivo. cbd bound to insoluble type i, ii, iii and iv collagens in vitro, and to skin, aorta, tendon, kidney, trachea and corneal tissues containing various types of collagen fibrils or sheets. cbd bound to all kinds of collagen fibrils regardless of their diameters and also bound to sheet-forming collagen in the glomerular basal lamina or desce ...	2001	11913772
screening of selected basidiomycetes for inhibitory activity on clostridium histolyticum collagenase and human leukocyte elastase.	collagenase and elastase play a critical role in the degradation of connective tissue. aqueous and dichloromethane extracts of 15 basidiomycetes were screened for their ability to inhibit the activity of collagenase and elastase. collagenase (ec 3.4.24.3) was not inhibited by aqueous extracts, but by dichloromethane extracts in concentrations of 200 microg/ml. for seven extracts an ic(50) less than 200 microg/ml could be observed. elastase (3.4.21.37) was inhibited by both aqueous and dichlorome ...	2002	11933148
bacterial protease inhibitors.	serine-, cysteine-, and metalloproteases are widely spread in many pathogenic bacteria, where they play critical functions related to colonization and evasion of host immune defenses, acquisition of nutrients for growth and proliferation, facilitation of dissemination, or tissue damage during infection. since all the antibiotics used clinically at the moment share a common mechanism of action, acting as inhibitors of the bacterial cell wall biosynthesis or affecting protein synthesis on ribosome ...	2002	12111749
protease inhibitors: synthesis of matrix metalloproteinase and bacterial collagenase inhibitors incorporating 5-amino-2-mercapto-1,3,4-thiadiazole zinc binding functions.	matrix metalloproteinase (mmp)/bacterial collagenase inhibitors incorporating 5-amino-2-mercapto-1,3,4-thiadiazole zinc binding functions are reported. a series of compounds was prepared by reaction of arylsulfonyl isocyanates or arylsulfonyl halides with phenylalanyl-alanine, followed by coupling with 5-amino-2-mercapto-1,3,4-thiadiazole in the presence of carbodiimides. these new compounds were assayed as inhibitors of human mmp-1, mmp-2, mmp-8 and mmp-9, and of the collagenase isolated from t ...	2002	12217351
free fatty acids inhibit the activity of clostridium histolyticum collagenase and human neutrophil elastase.	we investigated the ability of free fatty acids to inhibit the activity of clostridium histolyticum collagenase (ec 3.4.24.3) and human neutrophil elastase (ec 3.4.21.37). we determined the activity of collagenase by degradation of resorufin-labeled casein fluorimetrically. the determination of the elastase activity was performed by a spectrophotometric method using a 4-nitroanilide peptide substrate. we found that most of the tested fatty acids inhibited collagenase at concentrations between 50 ...	2002	12357383
mucosal and invading bacteria in patients with inflammatory bowel disease compared with controls.	endogenous intestinal bacteria and/or specific bacterial pathogens are suspected of being involved in the pathogenesis of inflammatory bowel diseases (ibd). the aim of this study was to investigate ibd tissues for different bacterial population groups harbouring the mucosal surface and/or invading the mucosa.	2002	12374228
pcr detection of clostridium chauvoei in pure cultures and in formalin-fixed, paraffin-embedded tissues.	the polymerase chain reaction (pcr) was used to amplify specific segments of the 16s ribosomal rna gene of clostridium chauvoei, a major pathogen of ruminants. three sets of primers were used to produce amplicons of 159, 836 and 959 base pairs (bp), respectively. the pcr was evaluated by testing clinically important strains of clostridium, including 21 strains of c. chauvoei, five strains each of clostridium septicum and clostridium perfringens and two strains each of clostridium novyi, clostrid ...	2003	12458172
inhibition of collagenase and metalloproteinases by aloins and aloe gel.	the effects of aloe barbadensis gel and aloe gel constituents on the activity of microbial and human metalloproteinases have been investigated. clostridium histolyticum collagenase (chc) results dose-dependently inhibited by aloe gel and the activity-guided fractionation led to an active fraction enriched in phenolics and aloins. aloins have been shown to be able to bind and to inhibit chc reversibly and non-competitively. aloe gel and aloins are also effective inhibitors of stimulated granulocy ...	2003	12479983
a quantitative structure-activity relationship study on some matrix metalloproteinase and collagenase inhibitors.	a quantitative structure-activity relationship (qsar) study is made on some hydroxamic acid-based inhibitors of matrix metalloproteinases (mmps) and a bacterial collagenase, namely clostridium histolyticum collagenase (chc), that also belongs to an mmp family, m-31, using kier's valence molecular connectivity index (1)chi(v) of the substituents and electrotopological state (e-state) indices of some atoms. the results indicate that out of the four mmps (mmp-1, mmp-2, mmp-8, and mmp-9) studied, mm ...	2003	12517437
collagenase-induced changes in articular cartilage as detected by electron-microscopic stereology, quantitative polarized light microscopy and biochemical assays.	the purpose of this study was to compare the ability of electron-microscopic (em) stereology with quantitative polarized light microscopy (plm) and biochemical collagen (hydroxyproline) and crosslink (pyridinoline) analyses to detect changes in the superficial collagen network of bovine articular cartilage after digestion in vitro with purified bacterial (clostridium histolyticum) collagenase (30 u/ml) for 24 and 48 h. collagen volume (v(v)) and surface (s(v)) densities of the uppermost third of ...	2002	12566629
a bacterial collagen-binding domain with novel calcium-binding motif controls domain orientation.	the crystal structure of a collagen-binding domain (cbd) with an n-terminal domain linker from clostridium histolyticum class i collagenase was determined at 1.00 a resolution in the absence of calcium (1nqj) and at 1.65 a resolution in the presence of calcium (1nqd). the mature enzyme is composed of four domains: a metalloprotease domain, a spacing domain and two cbds. a 12-residue-long linker is found at the n-terminus of each cbd. in the absence of calcium, the cbd reveals a beta-sheet sandwi ...	2003	12682007
protease inhibitors: synthesis of bacterial collagenase and matrix metalloproteinase inhibitors incorporating arylsulfonylureido and 5-dibenzo-suberenyl/suberyl moieties.	novel matrix metalloproteinase (mmp)/bacterial collagenase inhibitors are reported, considering the sulfonylated amino acid hydroxamates as lead molecules. a series of compounds was prepared by reaction of arylsulfonyl isocyanates with n-(5h-dibenzo[a,d]cyclohepten-5-yl)- and n-(10,11-dihydro-5h-dibenzo[a,d]cyclohepten-5-yl) methyl glycocolate, respectively, followed by the conversion of the coome to the carboxylate/hydroxamate moieties. the corresponding derivatives with methylene and ethylene ...	2003	12713832
a comparative qsar study on carbonic anhydrase and matrix metalloproteinase inhibition by sulfonylated amino acid hydroxamates.	a quantitative structure-activity relationship (qsar) study is made on the inhibition of a few isozymes of carbonic anhydrase (ca) and some matrix metalloproteinases (mmps), both zinc containing families of enzymes, by sulfonylated amino acid hydroxamates. for both enzymes, the inhibition potency of the hydroxamates is found to be well correlated with kier's first-order valence molecular connectivity index 1chi(v) of the molecule and electrotopological state indices of some atoms. from the resul ...	2003	12751815
a quantitative structure-activity relationship study on clostridium histolyticum collagenase inhibitors: roles of electrotopological state indices.	a quantitative structure-activity relationship (qsar) study has been made on eight different series of clostridium histolyticum collegenase (chc) inhibitors. these series are comprised of four different groups of sulfonylated amino acids and their corresponding hydroxamates. in each series, the inhibition potency of the compounds has been found to be significantly correlated with the electrotopological state (e-state) indices of nitrogen and sulfur atoms of the sulfonylated amino group in the mo ...	2003	12818668
a cysteine-activated proteinase of clostridium histolyticum.		1952	13000153
the partial purification of the lethal toxin of clostridium histolyticum.		1952	13006410